ANIMAL AND ANALYTICAL CORE (B) PROJECT SUMMARY The primary goal and function of Core B is to provide the appropriate animals and analytical assays required by projects in the Program Project. The objectives of Core B are to serve three basic support functions. First, Core B is responsible for maintaining the rodent colonies used in each Project. Second, Core B is responsible for generating the new collecting duct and vascular endothelial-specific knockout (KO) mice used in each Project. Third, Core B will provide analytical expertise and genotyping services. Core B will be a joint effort between the University of Utah (Dr. Donald Kohan) and the University of Alabama at Birmingham (UAB) (Dr. Jennifer Pollock). This arrangement will allow sharing of expertise in developing and breeding novel rodent models to provide the highest degree of quality control and allow consistency in animal models and analytical services across all of the individual projects of the Program Project. Aim 1 is to provide ETB-receptor deficient rats, Bmal1 KO rats, global P2X7 KO mice, global P2Y2 KO mice, LC-1 mice, and mice with collecting duct- specific KO of Bmal1, NOS1, ET-1 or ETB. This includes setting up the appropriate breeding pairs, assigning unique identifiers to each animal, tail clipping, genotyping, maintaining husbandry and genotyping records, and ensuring that a sufficient number of animals continue to be available for Projects 1, 2, and 3. Furthermore, it will include coordinating shipping of animals from Utah to UAB (Projects 1 and 3), and UAB to University of Texas Health Science Center at San Antonio (UTHSCSA) (Project 3). Aim 2 is to generate and provide the mice with inducible collecting duct-specific KO (polaris, polycystin-1, polycystin-2, or HDAC1), collecting duct- specific KO (NOS3, NOS1/NOS3), and vascular endothelial cell-specific KO (ET-1, ETB Bmal1). This activity includes breeding floxed polaris, floxed PC1, floxed PC-2 or floxed HDAC1 mice with HoxB7-rtTA and LC-1 mice to create inducible, collecting duct specific KO mice. Floxed (loxP-flanked) NOS3 or Bmal1 mice will be bred with AQP2-Cre mice to generate collecting duct specific KO of NOS3 or Bmal1, respectively. Tie2-Cre mice will be bred with floxed ET-1, ETB or floxed Bmal1 mice to generate vascular endothelial cell-specific KO of ET-1, ETB or Bmal1, respectively. These activities will occur at the University of Utah and UAB and will provide sufficient animals for all Projects. This includes setting up breeding pairs, assigning unique identifiers to each animal, tail clipping, genotyping, and maintaining husbandry and genotyping records. Aim 3 is to provide analytical expertise and genotyping services for of the gene-targeted animal models outlined in this PPG require genotyping. Core B will assist all Projects with analysis of genotyping results; in addition, given Dr. Kohan's expertise with confirming the degree and site(s) of gene targeting, Core B will assist all Projects with analysis of experiments characterizing the degree and sites of KO of the targeted allele(s). Aim 4 is to provide quality control and quantity purchasing for all mouse and rat studies for consistent and accurate results by monitoring and integrating procedures performed at the University of Utah, UAB, and the UTHSCSA.